• YAKHAK HOEJI
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• v.39 no.2
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• pp.161-168
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• 1995

Brain dopamine systems play a central role in the control of movement, hormone release, and many complex behavior. The action of dopamine at its synapse is terminated predominately by high affinity reuptake into presynaptic terminals by dopamine transporter (DAT). The dopamine transporter(DAT) is membrane protein localized to dopamine-containing nerve terminals and closely related with cocaine abuse, Parkinsonism, and schizophrenia. In present study, the recombinant plasmid pRc/CMV-DAT, constructed by subcloning of a cDNA encoding a bovine DAT into eukaryotic expression vector pRc/CMV, was stably transfected into CV-1 cells(monkey kidney cell line). The DAT activities in the cell lines selected by Geneticin$^{R}$ were determined by measuring the uptake of $[^3H]$-dopamine. The transfected cell lines showed 30-50 fold higher activities than untransfected CV-1 cell line, and this result implies that DAT is well expressed and localized in transfected cells. The transfected cells accumulated $[^3H]$-dopamine in a dose-dependent manner with a $K_{m}$ of 991.6nM. Even though high doses of norepinephrine, epinephrine, serotonin, and choline neurotransmitters inhibited the uptake of $[^3H]$-dopamine, DAT in transfected cell line was proven to be much more specific to dopamine. The psychotropic drugs such as GBR12909, CFT, normifensine, clomipramine, desipramine, and imipramine inhibited significantly the dopamine uptake in tissue culture cells stably transfected with DAT cDNA. Radioactive in situ hybridization was done to map the cellular localization of DAT mRNA-containing cells in the adult rat central nervous system. The strong hybridization signals were detected only in the substantia nigra pars compacta and ventral tegmental area. The restricted anatomical localization of DAT mRNA-containing cells confirms the DAT as a presynaptic marker of dopamine-containing cells in the rat brain.

• Molecules and Cells
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• v.40 no.7
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• pp.450-456
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• 2017

Mammalian physiology and behavior are regulated by an internal time-keeping system, referred to as circadian rhythm. The circadian timing system has a hierarchical organization composed of the master clock in the suprachiasmatic nucleus (SCN) and local clocks in extra-SCN brain regions and peripheral organs. The circadian clock molecular mechanism involves a network of transcription-translation feedback loops. In addition to the clinical association between circadian rhythm disruption and mood disorders, recent studies have suggested a molecular link between mood regulation and circadian rhythm. Specifically, genetic deletion of the circadian nuclear receptor Rev-$erb{\alpha}$ induces mania-like behavior caused by increased midbrain dopaminergic (DAergic) tone at dusk. The association between circadian rhythm and emotion-related behaviors can be applied to pathological conditions, including neurodegenerative diseases. In Parkinson's disease (PD), DAergic neurons in the substantia nigra pars compacta progressively degenerate leading to motor dysfunction. Patients with PD also exhibit non-motor symptoms, including sleep disorder and neuropsychiatric disorders. Thus, it is important to understand the mechanisms that link the molecular circadian clock and brain machinery in the regulation of emotional behaviors and related midbrain DAergic neuronal circuits in healthy and pathological states. This review summarizes the current literature regarding the association between circadian rhythm and mood regulation from a chronobiological perspective, and may provide insight into therapeutic approaches to target psychiatric symptoms in neurodegenerative diseases involving circadian rhythm dysfunction.

• Safety and Health at Work
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• v.7 no.1
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• pp.6-11
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• 2016

Background: A permit to work (PTW) is a formal written system to control certain types of work which are identified as potentially hazardous. However, human error in PTW processes can lead to an accident. Methods: This cross-sectional, descriptive study was conducted to estimate the probability of human errors in PTWprocesses in a chemical plant in Iran. In the first stage, through interviewing the personnel and studying the procedure in the plant, the PTW process was analyzed using the hierarchical task analysis technique. In doing so, PTWwas considered as a goal and detailed tasks to achieve the goal were analyzed. In the next step, the standardized plant analysis risk-human (SPAR-H) reliability analysis method was applied for estimation of human error probability. Results: The mean probability of human error in the PTW system was estimated to be 0.11. The highest probability of human error in the PTW process was related to flammable gas testing (50.7%). Conclusion: The SPAR-H method applied in this study could analyze and quantify the potential human errors and extract the required measures for reducing the error probabilities in PTW system. Some suggestions to reduce the likelihood of errors, especially in the field of modifying the performance shaping factors and dependencies among tasks are provided.

• Korean Journal of Veterinary Research
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• v.42 no.1
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• pp.7-20
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• 2002

This study deals with the distribution and morphological study of the neurons with the neuropeptide Y (NPY) in developing chick brain. The developing brains of Korean native chicks at embryonic days 8 (E8), E10, E12, E14, E16, E18, and E20 were used. The chicks were perfused with 4% paraformaldehyde through the left ventricle and aorta. The brains were removed and transferred into 30% sucrose, and then cut in a cryostat into $60{\mu}m$ in thickness. The sections were immunostained with free-floating and avidin-biotin complex (ABC) methods. The numerous neurons with NPY were first observed in nucleus rotundus of diencephalon at E8. In particular, neurons in nucleus rotundus had the well-developed processes. At E12, the neurons with NPY were distributed widely in diencephalon; nucleus septalis lateralis, medialis, nucleus magnocelluaris preopticus dorsalis, medialis, ventralis, nucleus preopticus medialis, nucleus dorsolateralis anterior thalami pars magnocellularis, and nucleus paraventricularis magnocellularis (paraventricular nucleus) except nucleus rotundus. From E12 to E20, positive neurons were oval-shaped, changed gradually into the spherical- and multipolar-shaped. The shapes of processes were also omnidirectional and the number of those were less than in telencephalon. As the chicks developed, the morphology of neurons with NPY showed the tendency to increase in their sizes and numbers.

• Parasites, Hosts and Diseases
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• v.38 no.3
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• pp.151-158
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• 2000

Pneumocystis carinii is a major opportunistic pathogen which has been found in the lungs of a wide variety of mammalian host species, and the fact suggests the possibility of intraspecific variation. Until now, P. carinii from different mammalian species are differentiated as subspecies, and the rats are known to be infected by two subspecies. The present study investigated genetic heterogeneity of P. carinii isolates from two strains of rats in Korea and China by molecular karyotyping, RFLP and sequencing analysis. Karyotypes of P. carinii were grouped into three, two from two strains of rats In Korea and one from rats in China. However RFLP of PCR product of ribosomal and MSG gene of the P. carinii isolates showed same pattern. The sequence homology rates of ${\alpha}-tubulin$ DNA of the P. carinii isolates were 96% in Seoul Wistar rats, 93% in Seoul Sprague-Dawley rats, and 85% in Chinese Sprague-Dawley rats. The present finding confirmed that P. carinii from rats in Korea are grouped into two karyotype strains which are different from that of P. carinii from rats in China. The Chinese isolate shows a little different sequences of ${\alpha}-tubulin$ DNA.

• Biomolecules & Therapeutics
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• v.27 no.2
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• pp.178-184
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• 2019

Parkinson's disease is a neurodegenerative disease characterized by the progressive loss of dopaminergic neurons within the substantia nigra pars compacta. In the present study, we investigated whether ${\beta}-Lapachone$ (${\beta}-LAP$), a natural naphthoquinone compound isolated from the lapacho tree (Tabebuia avellanedae), elicits neuroprotective effects in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced Parkinson's disease mouse model. ${\beta}-LAP$ reduced the tyrosine hydroxylase (TH)-immunoreactive fiber loss induced by MPTP in the dorsolateral striatum, and alleviated motor dysfunction as determined by the rotarod test. In addition, ${\beta}-LAP$ protected against MPTP-induced loss of TH positive neurons, and upregulated B-cell lymphoma 2 protein (Bcl-2) expression in the substantia nigra. Based on previous reports on the neuroprotective role of nuclear factor-E2-related factor-2 (Nrf2) in neurodegenerative diseases, we investigated whether ${\beta}-LAP$ induces upregulation of the Nrf2-hemeoxygenae-1 (HO-1) signaling pathway molecules in MPTP-injected mouse brains. Western blot and immunohistochemical analyses indicated that ${\beta}-LAP$ increased HO-1 expression in glial fibrillary acidic protein-positive astrocytes. Moreover, ${\beta}-LAP$ increased the nuclear translocation and DNA binding activity of Nrf2, and the phosphorylation of upstream adenosine monophosphate-activated protein kinase (AMPK). ${\beta}-LAP$ also increased the localization of p-AMPK and Nrf2 in astrocytes. Collectively, our data suggest that ${\beta}-LAP$ exerts neuroprotective effect in MPTP-injected mice by upregulating the p-AMPK/Nrf2/HO-1 signaling pathways in astrocytes.

• The Korea Journal of Herbology
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• v.28 no.5
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• pp.21-28
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• 2013

Objectives : The aim of this study was to investigate the neuroprotective effects and mechanisms of Cyperi Rhizoma extracts (CRE) using in vitro and in vivo models of Parkinson's disease (PD). Methods : We evaluated the neuroprotective effect of CRE against 1-methyl-4-phenylpyridinium (MPP+) toxicity using tyrosine hydroxylase immunohistochemistry (IHC) in primary rat mesencephalic dopaminergic neurons. In addition, the effect of CRE was evaluated in mice PD model induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). For evaluations, C57bl/6 mice were orally treated with CRE 50 mg/kg for 5 days and were injected intraperitoneally with MPTP (20 mg/kg) at 2 h intervals on the last day. To identify the CRE affects on MPTP-induced neuronal loss of dopaminergic neurons in substantia nigra pars compacta (SNpc) and striatum of mice, the behavioral tests and IHC analysis were carried out. Also, we conducted nitric oxide (NO) and tumor necrosis factor-alpha (TNF-${\alpha}$) assay in dopaminergic neurons and IHC using glial markers in SNpc of mice to assess the anti-inflammation effects. Results : In primary mesencephalic culture system, CRE protected dopaminergic cells against $10{\mu}M$ MPP+-induced toxicity at 0.2 and $1.0{\mu}g/mL$. In the behavior tests, CRE treated group showed improved motor deteriorations than those in the MPTP only treated group. CRE significantly protected striatal dopaminergic damage from MPTP-induced neurotoxicity in mice. Moreover, CRE inhibited productions of NO and TNF-${\alpha}$ in dopaminergic culture system and activation of astrocyte and microglia in SNpc of the mice. Conclusion : We concluded that CRE shows anti-parkinsonian effect by protecting dopaminergic neurons against MPP+/MPTP toxicities through anti-inflammatory actions.

• The Journal of Internal Korean Medicine
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• v.43 no.5
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• pp.1018-1028
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• 2022

Objectives: Parkinson's disease is characterized by progressive, irreversible damage to dopamine neurons in the substantia nigra pars compacta, as well as motor and non-motor symptoms. This disease currently has no dependable disease-modifying treatment. In this paper, we describe the treatment of a 67-year-old female with Parkinson's disease using acupuncture and exercise therapy. Case Presentation: Clinical symptoms and the United Kingdom Parkinson's Disease Society Brain Bank Diagnostic Criteria were used to diagnose the patient with Parkinson's disease. Over a 12-week period, the patient visited a Korean medicine hospital 18 times and was treated with acupuncture and exercise therapy in addition to anti-Parkinson's drugs. Before and after treatment, clinical examinations were performed using tools such as the Unified Parkinson's Disease Rating Scale, Fall Efficacy Scale, Parkinson's Disease Questionnaire, Berg Balance Scale, and Non-Motor Symptoms Scale. Furthermore, functional near-infrared spectroscopy was used to assess cortical hemodynamics. All clinical examination results improved after 12 weeks of intervention. In particular, improvements on the Total Unified Parkinson's Disease Rating Scale and Part III of this scale demonstrated large, clinically important differences. Conclusion: This case suggests that combining acupuncture and exercise therapy could produce an effective treatment for Parkinson's disease patients.

• Korean Journal of Acupuncture
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• v.39 no.2
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• pp.34-42
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• 2022

Objectives : Parkinson's disease (PD) is a neurodegenerative disease caused by dopaminergic neuronal death in the substantia nigra pars compacta. PD is known to be linked with mitochondrial dysfunction and increased oxidative stress. In this study, anti-cytotoxic and anti-oxidative effect of 12 herbal formulae were compared. Methods : According to experts' advice, 12 types of herbal formulae (Gamisoyosan, Galgeuntang, Galgeunhaegitang, Banhabaekchoolcheonmatang, Bojungikgitang, Boheotang, Sihogyejitang, Sihosogantang, Sihocheonggantang, Ojeoksan, Cheongsanggyeontongtang and Palmultang) were selected from 56 types of herbal formulae covered by the National Health Insurance Service in Korea. To detect anti-oxidative effect, 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay was performed, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to detect anti-cytotoxic effect of 12 herbal formulae using SH-SY5Y human neuroblastoma cells. Results : In DPPH assay, anti-oxidant activity was increased in a dose-dependent manner and half maximal inhibitory concentration was highest in the order of Galgeuntang, Gamisoyosan, Galgeunhaegitang, Ojeoksan, Palmultang, Sihogyejitang, Sihosogantang, Cheongsanggyeontongtang, Sihocheonggantang, Bojungikgitang, Boheotang and Banhabaekchoolcheonmatang. In MTT assay, concentration of 80% cell survival was highest in the order of Sihosogantang, Cheongsanggyeontongtang, Sihocheonggantang, Sihogyejitang, Bojungikgitang, Galgeuntang, Ojeoksan, Boheotang, Palmultang, Galgeunhaegitang, Banhabaekchoolcheonmatang and Gamisoyosan. Formulae with more than 50% DPPH radical scavenging activity at concentrations for 80% cell survival were Sihosogantang, Cheongsanggyeontongtang, Sihogyejitang, Galgeuntang and Sihocheonggantang. Conclusions : Sihosogantang, Cheongsanggyeontongtang, Sihogyejitang, Galgeuntang and Sihocheonggantang extracts can be candidate medicines for PD, but the effect should be validated in PD models.

• Journal of Korean Neurosurgical Society
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• v.30 no.6
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• pp.688-698
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• 2001

Objectives : Parkinson's disease is a well-known neurodegenerative disease characterized by dopaminergic cell death in the substantia nigra. The reactive gliosis by activated astrocytes and microglias is no more regarded as a simple sequel of neuronal cell death. Microglial activation takes place in a stereotypic pattern with graded morphologic and functional(resting, activated and phagocytic) changes. In Parkinson's disease animal model, the degree of microglial activation along the nigro-striatal dopaminergic tract has not been studied intensively. The purpose of this study was to elucidate the characteristics of microglial reaction and to grade its degree of activation at substantia nigra and corpus striatum using 6-hydroxydopamine induced rat model of Parkinson's disease. Methods : Using Sprague-Dawley rat, parkinsonian model was made by 6-hydroxydopamine(OHDA) induced destruction of medial and lateral substantia nigra(SN). The rat was sacrificed 3-, 5-, 7-, 14- and 21-day-after operation. For control group, we injected saline with same manner and sacrificed 3-day after operation. With immunohistochemistry, we examined dopaminergic neuronal cells and microglial expression using tyrosine hydroxylase (TH) and OX-42 antibodies, respectively. Also we performed in situ hybridization for osteopontin, a possible marker of subset in activated microglia. Results : 1) In lesioned side of substantia nigra and corpus striatum, the TH immunoreactivity was markedly decreased in whole experimental groups. 2) Using optical densitometry, microglia induced immunoreactivity of OX-42 was counted at SN and corpus striatum. At SN, it was increased significantly on the lesioned side in control and all time-dependent experimental groups. At striatum, it was increased significantly in post lesion 3-day group only(p <0.05). Compared to control group, immunoreactivity of OX-42 on lesioned side was increased in groups, except post lesion 21-day group, at SN. Only post lesion 3-day group showed significance at striatum(p <0.05). Compared to SN region, immunoreactivity of OX-42 was much weaker in striatum. 3) Microscopically, the microglias showed typically different activation pattern. At SN, numerous phagocytic microglias were found at pars compacta and reticularis of lesion side. At striatum, no phagocytic form was found and the intensity of staining was much weaker. 4) At SN, the immunoreactivity of osteopontin showed definite laterality and it was markedly increased at pars compacta of lesion side with relatively short duration time. At striatum, however, it was not detected by in situ hybridization technique. Conclusion : The nigral 6-OHDA induced rat model of Parkinson's disease revealed several characteristic patterns of microglial reaction. At SN, microglias was activated shortly after direct neuronal damage and maintained for about three weeks. In contrast, despite of sufficient dopaminergic insufficiency at striatum, activation of microglias was trivial, and distinguished 3 day later. Antegrade slow neuronal degeneration is major pathophysiology in striatal dopaminergic deficiency. So, the acuteness of neuronal damage and consequential degree of neuronal degeneration may be important factor for microglial activation in neurodegenerative diseases such as Parkinson's disease. Additionally, osteopontin may be a possible marker for several subsets of activated microglia, possibly the phagocytic form.